地中海实蝇幼虫分子鉴定

通过对从秘鲁进口的葡萄中截获实蝇类幼虫进行ITS区和线粒体COⅠ,COⅡ,COⅢ、ND5基因序列的扩增和测序,并与GenBank中对应的序列进行比对,结果表明,截获样品ITS区序列和地中海实蝇Ceratitis capitata(Wiedemann)同源性为95·16%(其中ITS1为99·52%,ITS2为86·2%),线粒体COⅠ,COⅡ,COⅢ,ND5基因序列和地中海实蝇C.capitata同源性为100%,99·9%,99·5%,99·8%;基于COⅠ序列构建的系统发育树中,幼虫样品和地中海实蝇最为接近。根据序列分析和系统发育关系分析的结果,将截获的实蝇类幼虫鉴定为地中海实蝇C.capitata。     

地中海实蝇地理种群遗传分化研究

以mtDNA的CO Ⅰ基因的部分序列作为分子标记,研究了来自安哥拉、黎巴嫩、刚果共和国、尼日利亚、贝宁、多哥、加纳、科特迪瓦、南非、埃及、法国、约旦、秘鲁、智利、美国佛罗里达和美国夏威夷等16个国家和地区共75个个体的地中海实蝇Ceratitis capitaza地理种群序列特征和单倍型特征,并分析了各种群间的遗传分化水平和进化关系,结果表明,在供试的样品中发现单倍型的数目是29,其中有5种为共享倍型.地中海实蝇各地理种群的遗传多态性水平差异较大,总体遗传距离D等于0.007.非洲南撒哈拉区(安哥拉、刚果、尼日利亚、贝宁、多哥、加纳、科特迪瓦)的种群的单倍型数、单倍型百分比和群内遗传距离等遗传多态性参数值都高于其它供试地区,多样性明显较其他地区丰富.     

中国实蝇检疫研究概况

实蝇（fruit fly）是为害水果和蔬菜的有害生物,这类害虫尤其是在果蔬的国际贸易中受到关注,多将其列为检疫性有害生物而置于有关的法规中。本文从应用于植物检疫的角度,着重就实蝇的监测和有关生物学的研究、实蝇鉴定技术研究、实蝇除害处理研究、建立实蝇非疫区、果园实蝇防治研究以及我国在水果国际双边贸易中的实蝇问题等多个方面的内容收集和介绍有关研究概况,对我国就以实蝇为内容的研究提供相关信息。     

地中海实蝇及其近缘种基因芯片检测研究

本研究选择线粒体DNA (mtDNA) 细胞色素氧化酶Ⅰ基因（COⅠ）为分子标记基因,以双翅目实蝇科昆虫DNA序列为目标,建立了我国进境植物检疫害虫地中海实蝇Ceratitis capitata、芒果小条实蝇C. cosyra和纳塔尔小条实蝇C. rosa等生物芯片检测方法。地中海实蝇及其近缘种检测芯片由检测探针（实蝇科通用探针1条,小条实蝇属通用探针1条,地中海实蝇、芒果小条实蝇和纳塔尔小条实蝇近缘种探针2条和种特异探针4条）、质控探针（定位点探针、阳性质控、阴性质控和空白对照探针各1条）组成。芯片检测结果表明,检测探针特异性强,能实现上述3种实蝇的种类快速区分和准确鉴定; 检测方法稳定性好,地中海实蝇不同虫态（卵、幼虫、蛹和成虫）和不同地理种群检测结果完全一致。地中海实蝇生物芯片检测技术将为我国进口果蔬中检疫性实蝇快速筛查和种类鉴定提供检测方法,同时,还可应用到其他属的实蝇以及相关害虫的检疫中,为有害生物的快速鉴定提供了新方法。     

橘小实蝇快速检疫鉴定方法

采用形态学观察与PCR技术相结合的方法,针对台湾水果中截获的可疑橘小实蝇Bactroceradorsalis(Hendel)幼虫和蛹进行快速鉴定。采用异硫氰酸胍方法快速提取昆虫基因组DNA,并根据特异性引物,采用PCR技术从截获的可疑橘小实蝇幼虫和蛹样本中均扩增得到224 bp的特异性条带,经测序比较发现,扩增产物序列与橘小实蝇目的基因序列完全相同,由此证明可疑样本为橘小实蝇的幼虫和蛹。该方法解决了橘小实蝇幼虫和蛹等未成熟虫态的快速检疫难题,大大缩短了检测周期,值得口岸检验检疫借鉴应用。     

凤实蝇属的分类厘订 (双翅目: 实蝇科: 实蝇亚科)

在实蝇亚科Trypetinae内,由于凤实蝇属Felderimyia Hendel与邻实蝇属Ptilona van der Wulp在诸多形态特征和雌性外生殖器构造方面十分近似,因此此属应归隶于刺脉实蝇族Acanthonevrini。对凤实蝇属进行了重新界定,详细描述中国1新纪录种：黑翅凤实蝇F.fuscipennis Hendel,并提供分种检索表和特征图。     

实蝇类昆虫的引诱剂和诱捕器

实蝇科昆虫的许多种类是重要的入侵物种, 在国际植物检疫领域具有重要地位。联合国粮农组织（FAO）和《国际植物保护公约（IPPC）》积极倡导国际合作实施对实蝇类害虫的有效控制, 不断完善和出版实蝇控制技术手册并制定相关的国际植物检疫标准。引诱剂和诱捕器是实蝇类害虫监测、调查和防治中最重要的手段之一, 被广泛采用。国际上普遍认可的实蝇诱剂主要包括蛋白质诱剂（PA)、甲基丁香酚（ME）、诱蝇酮（CUE）、乙酸铵诱剂（AA）、乙酸铵盐诱剂（AS）、TML 诱剂、己酸丁酯诱剂（BuH）、吡嗪诱剂（MVP）等, 我国也研发了大量植物型和食物型诱剂。国际上常用实蝇诱捕器有20多种, 依据捕获实蝇的不同方式分为干型诱捕器、湿型诱捕器和干湿综合型诱捕器3类。同时, 我国利用各种塑料瓶研制了多种多样的诱捕器。本文对国际、国内实蝇诱剂和诱捕器等研究和利用情况进行了总结, 提出我国在建设外向型水果非疫区的实蝇监测中应使用国际标准的诱剂和诱捕器, 以便获得国际认可并促进我国优势农产品顺利进入国际市场, 在研发诱剂和诱捕器时做到标准化并及时申请专利保护、推向国际市场以便获得国际有关标准的采用和认可。     

中国寡鬃实蝇属记述：双翅目：实蝇科

中国寡鬃实蝇属 Dacus Fabrcius目前已知6亚属31种。本文记述其中的5个新种:锈红寡鬃实蝇D.(Bactrocera)rubiginus sp.nov.,海南寡鬃实蝇D.(Didacus)hainanus sp.nov.滇寡鬃实蝇D.(Asiadacus)dianensis sp.nov.,盾条寡鬃实蝇D.(Zeugodacus)adustus sp.nov.,狭腹寡鬃实蝇D.(Zeugodacus)stenomus sp.nov.;记载5个中国新纪录:实胫寡鬃实蝇D.(Bactrocera)correctus(Bezzi),短条寡鬃实蝇D.(Zeugodacus)abbreviatus Hardy,印度寡鬃实蝇D.(Zeugodacus) scutellaris(Bezzi),番石榴寡鬃实蝇D.(Hemigymnodacus)diversus Coq.,黑背寡鬃实蝇D.(Paratridacus)expandens melanius Hardy & Adachi;并列出种检索表。     

Determination of host status of citrus fruits against the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae)

Abstract  The Mediterranean fruit fly (Medfly), Ceratitis capitata (Wiedemann) is a pest of citrus in parts of Western Australia. Three citrus cultivars: Valencia oranges, Eureka lemons and Imperial mandarins, as well as non-citrus control fruits, were examined for attractiveness and suitability to Medfly in the field and in the laboratory using choice and no-choice experiments. Oranges were more susceptible to Medfly than mandarins and lemons. Punctures in the skin had a significant impact on the degree of infestation in both citrus and non-citrus control fruit. Artificial infestation and larval survivorship were used to investigate the suitability of each cultivar to Medfly under laboratory conditions. Oranges and mandarins were suitable for the development of Medfly, but lemons were a poor host. When each cultivar was in season, field cage trials demonstrated that infestation occurred in oranges and mandarins but not in lemons.     

Mitochondrial DNA variation in Spanish populations of Ceratitis capitata (Wiedemann) (Tephritidae) and the colonization process

Abstract: Mitochondrial DNA variation in the Mediterranean fruit fly Ceratitis capitata (Wiedemann, Diptera: Tephritidae) was studied in three natural populations from southern, central and eastern Spain by means of restriction fragment length polymorphisms using 22 restriction endonucleases. Nine different haplotypes were found based upon the restriction patterns of the seven polymorphic endonucleases, providing a measure of discrimination between populations (N_ST = 0.2462, F_ST = 0.154). The observed distribution of haplotypes, corroborated by a parsimonious unrooted tree, suggests an ancient origin for haplotype VII, and a first step in the colonization of the Iberian Peninsula and subsequently the northern and eastern Mediterranean basin, through the Straits of Gibraltar. No relationship can be established between the colonization process for Europe and America.     

Glycosylation and adhesiveness differentiate larval Ceratitis capitata tyrosinases

Larval Ceratitis capitata phenoloxidases (POs) from hemocytes, serum, integument, and fat body were analyzed. Two types of PO were recorded: the tyrosinase type found in hemocytes, serum, integument, and fat body and the laccase type found in integument. Tyrosinase from all larval tissues and integumental laccase as well, showed similarity in molecular weight (93 KDa), activation by Escherichia coli at 5 mM Ca²⁺, and reactivity to antibodies raised against serum tyrosinase. However, the enzymes differed with respect to their glycosylation and adhesiveness. The serum and integumental enzyme forms contain concanavalin A reacting material, whereas hemocyte and integumental tyrosinase(s) are adhesive. These differences in enzyme forms, although not influencing their substrate specificity, seem to give advantages to performing their function, i.e., the adhesive enzyme form facilitates the adherence to E. coli cell wall and hemocyte surface (unpublished data) while the glycosylated form facilitated the secretion into serum. © 1994 Wiley-Liss, Inc.     

The fruit fly PUB: a phagostimulation unit bioassay system to quantitatively measure ingestion of baits by individual flies

Abstract:  A bioassay to investigate quantitative phagostimulation and ingestion physiology of baits on individual fruit flies is presented. The study was undertaken using two fruit fly species: the Mediterranean fruit fly ( Ceratitis capitata ), a cosmopolitan insect pest, and the Ethiopian fruit fly ( Dacus ciliatus ), a quarantine insect in Israel. Our model bait suspension included spinosad as the toxic agent, and 1% yeast hydrolysate with 10% sucrose as phagostimulant. A preliminary toxicology study showed that the two fruit flies are highly sensitive to low concentrations of spinosad baited with this phagostimulant. The maximum concentration needed to kill 90% of the female flies was 4.2 and 8.5 p.p.m. for C. capitata and D. ciliatus , respectively. The bioassay was able to detect the ingestion of low volumes (e.g. 1  μ l) of tested solutions. The bioassay was also able to detect differences in intake of different concentrations of spinosad solutions and relate ingestion to fruit fly mortality. Additionally, the bioassay was sensitive enough to highlight differences in intake related to the physiological status of the fruit fly and fly species. The bioassay can also be used to follow ingestion kinetics of baits. We expect that this bioassay will contribute in the exploration of more efficient bait systems for fruit flies.     

Effects of egg load on finding and acceptance of host fruit in Ceratitis capitata flies

Abstract. In studies conducted on potted host trees in field cages and in the laboratory, we examined the influence of egg load on the finding and acceptance of high-ranking (kumquat) and lower-ranking (grapefruit) hosts for oviposition by wild-origin Mediterranean fruit fly females, Ceratitis capitata (Wiedemann). By prescribing the periods during which females had access to protein prior to testing, we generated four classes of females having progressively increasing egg loads but not differing in population origin, age, degree of protein hunger at testing, or amount of prior experience with host fruit (none). Egg load had no discernible effect on behaviour associated with finding either type of fruit but did have a significant effect on several behaviours associated with oviposition after alighting on fruit. Increasing egg load led to increasing propensity to engage in ovipositional-type behaviour on both kumquats and grapefruits. There was no evidence, however, to support a hypothesis that medflies would become less discriminating against grapefruits relative to kumquats as egg load increased. Relative to kumquats, grapefruits were accepted for oviposition by intermediate and high egg load females to a substantially greater degree in laboratory cages than on trees, suggesting that results of laboratory cage experiments on host discrimination by tephritid flies may poorly reflect differences in behavioural responses expressed under less constrained conditions.     

Investigation of native isolates of entomopathogenic fungi for the biological control of three citrus pests

The pathogenicity of 15 isolates of Beauveria bassiana (Balsamo) Vuillemin, five isolates of Metarhizium anisopliae (Metschnikoff) Sorokin and one isolate of M. flavoviride (Gams and Rozsypal) were tested under laboratory conditions against the subterranean life stages of the citrus pests, Ceratitis rosa Karsch, C. capitata Wiedemann (Diptera: Tephritidae) and Thaumatotibia leucotreta Meyrick. (Lepidoptera: Tortricidae). When these citrus pests were treated with a concentration of 1×10⁷ conidia mL^?1, fungal isolates had a significantly greater effect on the adults of C. rosa and C. capitata than they did on the puparia of these two fruit fly species. Further, C. rosa and C. capitata did not differ significantly in their response to entomopathogenic fungi when adult and pupal mycosis were considered. Depending on fungal isolate, the percentage of T. leucotreta adults which emerged from fungal treated sand ranged from 5 to 60% and the percentage of pupae with visible signs of mycosis ranged from 21 to 93%. The relative virulence of the four most promising fungal isolates, as well as the commercially available B. bassiana product, BroadBand® (Biological Control Products, South Africa), were compared against one another as log-probit regressions of mortality against T. leucotreta which exhibited a dose-dependent response. The estimated LC₅₀ values of the three most virulent B. bassiana isolates ranged from 6.8×10⁵ to 2.1×10⁶ conidia mL^?1, while those of the least pathogenic ranged from 1.6×10⁷ to 3.7×10⁷ conidia mL^?1.     

Detection of novel mitochondrial haplotype variants in populations of the Mediterranean fruit fly,Ceratitis capitata,from Tunisia,Israel and Morocco

The direct analysis of DNA sequence‐based mitochondrial haplotypes in individuals from populations of the Mediterranean fruit fly, Ceratitis capitata, from Tunisia, Morocco and Israel has revealed levels of genetic diversity that far exceed those previously described. A total of 33 distinct haplotypes were identified from 117 individuals representing seven populations from these countries. A network of haplotypes was constructed showing that we have captured essentially all of the sequence variations present in these populations for this gene. These results have important implications for future studies of the genetic structure of this and other closely related pest species.     

Synthesis and deposition of PCG-100, the main pupal cuticle glycoprotein of the medfly Ceratitis capitata

We have studied the developmental expression of the main pupal cuticle glycoprotein, PCG-100, in the Medfly Ceratitis capitata. A polyclonal antiserum was raised against this protein. Western blotting analysis showed that this glycoprotein is integument- and stage-specific. No PCG-100 or immunologically related polypeptides were detected in other tissues or instars. As studied by microinjection of [³⁵S]methionine in individual flies, in vivo synthesis and deposition of PCG-100 begins approximately 48 h after the onset of pupariation, shortly after the time of head eversion. Synthesis is maximal at 54–64 h, decreases at 72 h, and practically ceases in fully shaped 4-day-old pupae. The time required for PCG-100 deposition into the cuticle was found to be less than 10 min after its synthesis. This is the first time such in vivo analysis has been performed. © 1994 Wiley-Liss, Inc.